A survey of filamentous bacterial populations from foaming activated sludge plants in Eastern States of Australia

Of 129 activated sludge plants surveyed during 1988 in Queensland, New South Wales and Victoria, 66 had a foam problem during the sampling period. Filament identification revealed that Microthrix parvicella, Nocardia amarae and Nocardia pinensis were the most commonly found, followed by Eikelboom types 0092, 0914 and 0041/0675, although their relative frequencies varied from state to state.     

An examination of the mechanisms for stable foam formation in activated sludge systems

Screening pure cultures of 65 mycolic acid producing bacteria (Mycolata) isolated mainly from activated sludge with a laboratory based foaming test revealed that not all foamed under the conditions used. However, for most, the data were generally consistent with the flotation theory as an explanation for foaming. Thus a stable foam required three components, air bubbles, surfactants and hydrophobic cells. With non-hydrophobic cells, an unstable foam was generated, and in the absence of surfactants, cells formed a greasy surface scum. Addition of surfactant converted a scumming population into one forming a stable foam. The ability to generate a foam depended on a threshold cell number, which varied between individual isolates and reduced markedly in the presence of surfactant. Consequently, the concept of a universal threshold applicable to all foaming Mycolata is not supported by these data. The role of surfactants in foaming is poorly understood, but evidence is presented for the first time that surfactin synthesised by Bacillus subtilis may be important.     

Long-term analysis of a full-scale activated sludge wastewater treatment system exhibiting seasonal biological foaming

The seasonal accumulation of biological foam on the activated sludge system of the Urbana-Champaign Sanitary District Northeast (UCSD-NE) wastewater treatment plant was investigated over an 8-year period by statistical analyses including path analysis, multivariate regression, and principal component analysis. Results of these analyses suggested that variation in the activated sludge reactor temperature and the use of a stream bypassing the primary clarifier were the two main factors determining the observed temporal foam profile. Characterization of the primary clarifier influent and effluent suggested the involvement of high lipid loading rates from the bypass stream in foam accumulation. In light of these results, it is hypothesized that increasing temperatures and lipid loading rates are responsible for foam formation through the same mechanism: the foam-forming microbial population is specialized in consuming lipids, substrates classified as slowly degradable. When the temperature increases, the rate of lipid hydrolysis becomes sufficiently high for this population to become abundant, accumulate on the surfaces of the aeration basins, and cause biological foaming.     

利用剩余污泥制备泡沫灭火剂的试验研究

以武汉市某污水处理厂的剩余污泥为材料,在不同酸、碱浓度以及不同时间条件下对其进行水解,以制备含水解蛋白质、脂肪酸、糖类等可溶性有机物及无机物的混合液,并分析、测定了所制备混合液的蛋白质含量,探讨了影响剩余污泥水解的因素.试验结果表明,当采用NaOH水解剩余污泥时,所产水解液为膏状,不能过滤,因而不适合采用;而HCl可水解剩余污泥,且pH值为1.0时的水解效果最好,蛋白质含量高（59.2%）,但所制备的蛋白质混合液的发泡倍数相对较小（7.0倍）;CaO水解剩余污泥的效果较好,所制备蛋白质混合液的发泡倍数较高（7.6倍）,但所需时间较长,且蛋白质含量不高、CaO用量较大;将CaO（25%,相对于干污泥的投加量）与NaOH（5%,相对干污泥的投加量）混合使用的效果最好,不仅水解时间短,而且所得水解液可过滤、蛋白质含量高.经过测定同一蛋白质样品在不同水解时间下的灭火参数发现,水解时间越短则其发泡倍数越高.通过测定不同混合液与粘度、沉淀物、发泡倍数、90%火焰控制时间、灭火时间、抗烧时间等灭火指标的关系,分析了剩余污泥水解液的灭火机理.结果表明,剩余污泥水解液属于空气泡沫灭火剂,其灭火指标达到了国家标准（GA 219-1999）.此技术可实现剩余污泥的再利用,既避免了剩余污泥的二次污染,又可克服使用化学泡沫灭火剂时对环境的污染.     

The nature of activated sludge flocs

Previous work has suggested that the settling properties of sludge depend on the chemical and physical nature of the floc surface and that in at least one case the sludge volume index was directly related to the surface charge carried by the sludge particles. This relationship has now been confirmed for sludges from three other works. These sludges all had surface “slime-layers” which contained a polysaccharide composed of neutral sugars and glucuronic acid. In addition to this however, a lysozyme sensitive material was found which also contributed to the overall charge on the particles. This latter material was thought to originate from dead bacteria. Polyvalent cations were also found to be involved in the floc structure and also had an influence on the surface charge. The adsorption of cations by bacteria and biopolymers showed that there were two different types of adsorption behaviour for the activated sludge surface polymer depending on whether it was in the solid state or in solution. It was therefore suggested that different adsorption sites were available to the cations, carboxyl groups being proposed for soluble polymers and hydroxyl groups for the solid. The significance of this adsorption data to the removal of heavy metal ions by the activated sludge process is also discussed.     

A novel bioflocculant produced by Klebsiella sp. and its application to sludge dewatering

A bioflocculant‐producing bacterium, named N‐10, was isolated from activated sludge and was identified as Klebsiella sp. The bioflocculant (named MBF10) produced by Klebsiella sp. had a good flocculating capability and could achieve a flocculating rate of 86.5% to kaolin suspension at the dosage of 0.034 g/L. Compared with chemical flocculants, such as Al₂(SO₄)₃, polyaluminum chloride (PAC) and cationic polyacrylamide (PAM) at their optimal dosages, MBF10 showed a similar performance for sludge dewatering. After preconditioning, dry solids (DS) and specific resistance to filtration (SRF) of the sludge were 17.5% and 3.36 × 10¹² m/kg, respectively. The combined use of MBF10 and Al₂(SO₄)₃ resulted in optimum sludge dewatering properties, SRF reduced from 10.87 × 10¹² m/kg to 1.72 × 10¹² m/kg, and DS increased from 13.1% to 21.3%. Charge neutralisation and interparticle bridging were proposed as the reasons for the enhanced performance based upon the experimental observations.     

Stable Foams and Sludge Bulking: The Largest Remaining Problems (Abridged)

The separation of activated sludge in secondary settling tanks is the crucial step in biological wastewater treatment from the point of view of both final effluent quality and operation of the aeration plant. The sludge-separation problems of bulking and foaming are connected with an excessive growth of filamentous micro-organisms in the biocenosis of activated sludge.
This paper (a) describes the methods which are used for the quantification of activated-sludge separation problems, and (b) summarizes the design recommendations for bulking and foaming control.     

Progress and perspectives of sludge ozonation as a powerful pretreatment method for minimization of excess sludge production

The treatment and disposal of excess sludge represents a bottleneck in wastewater treatment plants (WWTP) worldwide, due to environmental, economic, social and legal factors. The ideal solution to the problem of sludge disposal is to combine sludge reduction with the removal of pollution at the source. This paper presents an overview of the potential of ozonation in sludge reduction. The full-scale application of ozonation in excess sludge reduction is presented. Improvements in the biodegradability of the ozonated sludge were confirmed. The introduction of ozonation into activated sludge did not significantly influence effluent quality but improved the settling properties of the sludge. An operation with a suitable sludge wasting ratio seems to be necessary to prevent accumulation of inorganic and inert particles for long-term operation. Sludge ozonation to reduce excess sludge production may be economical in WWTP which have high sludge disposal costs and operational problems such as sludge foaming and bulking. The recommended ozone dose ranges from 0.03 to 0.05 g O₃/g TSS, which is appropriate to achieve a balance between sludge reduction efficiency and cost. An effort to design and optimize an economic sludge reduction process is necessary.     

Ecological study of activated sludge settling property in the completely mixed system

The bulking problem associated with both nitrogen-rich and nitrogen-deficient activated sludges was thoroughly investigated under a DO sufficient condition in the completely-mixed continuous flow system. The reactor temperature was controlled at 27 ± 1°C and the pH was maintained at nearly equal to 7.0. The activated sludge cultures were acclimated under various organic loadings and their settling characteristics were determined by sludge volume index and zone settling velocity. The contents of sludge protein and carbohydrate along with the cell surface charge were examined for understanding the correlation between sludge composition and sludge settleability. Also, the major types of long-length filaments existed in the bulky activated sludge were identified. Moreover, the effect of sludge settleability on effluent quality was determined.     

Challenges in determining causation in structure-function studies using molecular biological techniques

The use of molecular biological techniques for determining the levels and types of different microbial populations in bioreactors has led to the emergence of the microbial community ‘structure-function’ paradigm that is often used in research. Typically, lab- or full-scale systems are monitored for the relevant parameters, and these parameters are related to the changes in microbial populations. Research in activated sludge phenomena, such as filamentous bulking, filamentous foaming, nitrogen removal, and phosphorus removal, are replete with many examples of this ‘structure-function’ paradigm, most commonly those that involve 16S rRNA gene-based analysis of the microbial populations. In many cases, such studies assume a causal microbial population (e.g., a species that causes bulking or foaming), or conclude in identifying a causal population. However, assigning cause to specific organisms and populations is problematic in a complex environment such as wastewater bioreactors. The Koch-Henle postulates, the gold standard in evaluating causation of disease, have limitations when applied to systems with mixed microbial communities with complex interactions, particularly if pure cultures are not available. Molecular techniques that allow specific identification and quantification of organisms have been used by researchers to overcome the limitations of culture-based techniques, and at the same time, raised new questions on the applicability of causation postulates in environmental systems. In this paper, various causation criteria improving on the Koch-Henle postulates are presented. Complicating issues in assigning cause in wastewater bioreactors are identified. Approaches for determining cause-effect relationships are illustrated using 16S rDNA-based investigations of filaments that cause bulking and foaming in activated sludge. The hope is that a causation framework that accounts for the assumptions in molecular studies, as applied to wastewater treatment research, will lead to improved experimental design and analysis of data.     

Ultrasonic enhancement of waste activated sludge hydrolysis and volatile fatty acids accumulation at pH 10.0

Volatile fatty acids (VFA), the preferred carbon source for biological nutrients removal, can be produced by waste activated sludge (WAS) anaerobic fermentation. However, because the rate of VFA accumulation is limited by that of WAS hydrolysis and VFA is always consumed by methanogens at acidic or neutral pHs, the ultrasonic pretreatment which can accelerate the rate of WAS hydrolysis, and alkaline adjustment which can inhibit the activities of methanogens, were, therefore, used to improve WAS hydrolysis and VFA accumulation in this study. Experiment results showed that the combination of ultrasonic pretreatment and alkaline adjustment caused significant enhancements of WAS hydrolysis and VFA accumulation. The study of ultrasonic energy density effect revealed that energy density influenced not only the total VFA accumulation but also the percentage of individual VFA. The maximal VFA accumulation (3109.8 mg COD/L) occurred at ultrasonic energy density of 1.0 kW/L and fermentation time of 72 h, which was more than two times that without ultrasonic treatment (1275.0 mg COD/L). The analysis of VFA composition showed that the percentage of acetic acid ranked the first (more than 40%) and those of iso-valeric and propionic acids located at the second and third places, respectively. Thus, the suitable ultrasonic conditions combined with alkaline adjustment for VFA accumulation from WAS were ultrasonic energy density of 1.0 kW/L and fermentation time of 72 h. Also, the key enzymes related to VFA formation exhibited the highest activities at ultrasonic energy density of 1.0 kW/L, which resulted in the greatest VFA production during WAS fermentation at pH 10.0.     

Activated sludge deflocculation under temperature upshifts from 30 to 45 degrees C

Twelve independent batch experiments (<9h) with fresh municipal activated sludge were conducted to assess the occurrence and the mechanisms of deflocculation under a temperature shift from 30 to 45 degrees C. In each experiment, a transient reactor (2 L) was subjected to the temperature shift and a control reactor was operated at a constant temperature of 30 degrees C. The occurrence of deflocculation was demonstrated by the increase in turbidity and in the concentrations of biopolymers in the sludge supernatant from the transient reactor. The maximum levels of proteins in the supernatants ranged from 53 to 81 mg/L, for DNA from 34 to 36 mg/L, for humic compounds from 20 to 40 mg/L, and for carbohydrates from 21 to 31 mg/L. All the biopolymer concentrations in the control reactor remained below 5-10 mg/L. The release of biopolymers was accompanied by an increase in sludge supernatant conductivity (16-32% increase, up to 1.20 mS/cm), soluble chemical oxygen demand (from 129 to 440 mg/L), total suspended solids (>25 mg/L up to 128 mg/L), and a decrease in the mixed liquor volatile suspended solids (up to 11%). The temperature shift was also found to inhibit microbial metabolism by reducing the sludge biomass substrate removal capacity, as measured by oxygen-uptake rates. The temperature shift had a marginal effect causing sludge lysis (as an increase in beta-galactosidase activity) and had no significant impact on sludge viability (live/dead ratio of bacterial cells). It was concluded that sludge deflocculation under a temperature shift from 30 to 45 degrees C involves the solubilisation of extracellular polymeric substances from the flocs and likely also floc fragmentation. In addition, sludge deflocculation and the inhibition of microbial metabolism explain the poor treatment performance observed in previous continuous reactors under similar temperature shifts.     

污水处理中CASS工艺活性污泥培养与驯化

通过重庆市大渡口污水处理厂工程,重点介绍了工程调试中活性污泥培养与驯化的情况。实践表明,采用接种法培养驯化活性污泥20d就能完成污泥培养驯化,COD、BOD、SS的去除率分别为77.8%～88.3%、88.4%～95.9%、83.1%～92.1%,各项出水水质指标达到或优于设计目标。最后对培养驯化过程中出现的泡沫、进水BOD低营养源不足、滗水后期有微弱曝气和污泥回流、排泥等问题提出了相应的解决措施。     

Fate and removal of selected endocrine‐disrupting compounds in sewage using activated sludge treatment

The variation and fate of four endocrine‐disrupting compounds (EDCs) composed of 4‐n‐nonylphenol (4‐n‐NP), bisphenol A (BPA), 17β‐estradiol (E2) and 17α‐ethinylestradiol (EE2) were investigated along treatment units in a sewage treatment plant (STP), China with anaerobic, anoxic and aerobic activated sludge processes. The mean concentrations were 64.8 ng/L (E2), 115.3 ng/L (4‐n‐NP), 171.5 ng/L (EE2), and 920.7 ng/L (BPA) in the influents, and 22.8 ng/L (E2), 50.9 ng/L (4‐n‐NP), 49.9 ng/L (EE2), and 84.3 ng/L (BPA) in the effluents. The biological treatment was more effective in removing NP, BPA and E2 from the aqueous phase than the primary treatment, while the latter could effectively remove EE2. Their possible removal mechanisms during the biological treatment with activated sludge were further explored through spiked batch experiments under three oxygen‐supplying conditions (anaerobic, anoxic and aerobic). The batch experiments showed that 4‐n‐NP, E2 and BPA were removed from the aqueous phase through biodegradation. The combination of sludge sorption and biodegradation accounted for the removal of EE2. Anoxic activated sludge showed the most rapid degradation of 4‐n‐NP, while E2 could be removed most effectively by aerobic activated sludge, and sludge sorption had a remarkable effect on its removal within the initial 15 min of the experiments under three oxygen‐supplying conditions.     

Effects of sonication on bacteria viability in wastewater treatment plants evaluated by flow cytometry--fecal indicators, wastewater and activated sludge

The application of sonication to wastewater or sludge contributes to the dispersion of aggregates, the solubilisation of particulate matter with an increase in its biodegradability, the damage of microorganisms due to the loss of cellular membrane integrity. This research is aimed at investigating the effects of sonication at 20kHz frequency on viability of microorganisms present in raw wastewater and activated sludge taken from a municipal wastewater treatment plant, as well as pure strains of Escherichia coli and E. faecalis. Flow cytometry was applied for the identification and quantification of viable and dead bacteria free in the bulk liquid, after the fluorescent staining of cellular nucleic acids. The main results showed that: (i) cells of E. coli were highly sensitive to sonication, even at low specific ultrasonic energy (E(s)), and disintegration of a large amount of cells was observed; (ii) on the contrary E. faecalis were more resistant than E. coli, even if high levels of E(s) were applied; (iii) bacteria in raw wastewater exhibited a dynamic of viable and dead bacteria similar to E. coli; (iv) in activated sludge samples, low levels of E(s) produced a prevalent disaggregation of flocs releasing single cells in the bulk liquid, while disruption of bacteria was induced only by very high levels of E(s).     

Waste activated sludge hydrolysis and short-chain fatty acids accumulation under mesophilic and thermophilic conditions: Effect of pH

The effect of pH (4.0–11.0) on waste activated sludge (WAS) hydrolysis and short-chain fatty acids (SCFAs) accumulation under mesophilic and thermophilic conditions were investigated. The WAS hydrolysis increased markedly in thermophilic fermentation compared to mesophilic fermentation at any pH investigated. The hydrolysis at alkaline pHs (8.0–11.0) was greater than that at acidic pHs, but both of the acidic and alkaline hydrolysis was higher than that pH uncontrolled under either mesophilic or thermophilic conditions. No matter in mesophilic or thermophilic fermentation, the accumulation of SCFAs at alkaline pHs was greater than at acidic or uncontrolled pHs. The optimum SCFAs accumulation was 0.298 g COD/g volatile suspended solids (VSS) with mesophilic fermentation, and 0.368 with thermophilic fermentation, which was observed respectively at pH 9.0 and fermentation time 5 d and pH 8.0 and time 9 d. The maximum SCFAs productions reported in this study were much greater than that in the literature. The analysis of the SCFAs composition showed that acetic acid was the prevalent acid in the accumulated SCFAs at any pH investigated under both temperatures, followed by propionic acid and n-valeric acid. Nevertheless, during the entire mesophilic and thermophilic fermentation the activity of methanogens was inhibited severely at acid or alkaline pHs, and the highest methane concentration was obtained at pH 7.0 in most cases. The studies of carbon mass balance showed that during WAS fermentation the reduction of VSS decreased with the increase of pH, and the thermophilic VSS reduction was greater than the mesophilic one. Further investigation indicated that most of the reduced VSS was converted to soluble protein and carbohydrate and SCFAs in two fermentations systems, while little formed methane and carbon dioxide.     

Micro-profiles of activated sludge floc determined using microelectrodes

The microbial activity within activated sludge floc is a key factor in the performance of the activated sludge process. In this study, the microenvironment of activated sludge flocs from two wastewater treatment plants (Mill Creek Wastewater Treatment Plant and Muddy Creek Wastewater Treatment Plant, with aeration tank influent CODs of 60-120 and 15-35 mg/L, respectively) were studied by using microelectrodes. Due to microbial oxygen utilization, the aerobic region in the activated sludge floc was limited to the surface layer (0.1-0.2mm) of the sludge aggregate at the Mill Creek plant. The presence of an anoxic zone inside the sludge floc under aerobic conditions was confirmed in this study. When the dissolved oxygen (DO) in the bulk liquid was higher than 4.0mg/L, the anoxic zone inside the activated sludge floc disappeared, which is helpful for biodegradation. At the Muddy Creek plant, with its lower wastewater pollutant concentrations, the redox potential and DO inside the sludge aggregates were higher than those at the Mill Creek plant. The contaminant concentration in the bulk wastewater correlates with the oxygen utilization rate, which directly influences the oxygen penetration inside the activated sludge floc, and results in redox potential changes within the floc. The measured microprofiles revealed the continuous decrease of nitrate concentration inside the activated sludge floc, even though significant nitrification was observed in the bulk wastewater. The oxygen consumption and nitrification rate analyses reveal that the increase of ammonia flux under aerobic conditions correlates with nitrification. Due to the metabolic mechanisms of the microorganisms in activated sludge floc, which varies from one treatment plant to another, the oxygen flux inside the sludge floc changes accordingly.     

Optimization of PMA‐qPCR for Staphylococcus aureus and determination of viable bacteria in indoor air

Staphylococcus aureus may cause infections in humans from mild skin disorders to lethal pneumonia. Rapid and accurate monitoring of viable S. aureus is essential to characterize human exposure. This study evaluated quantitative PCR (qPCR) with propidium monoazide (PMA) to quantify S. aureus. The results showed comparable S. aureus counts between exclusively live cells and mixtures of live/dead cells by qPCR with 1.5 or 2.3 μg/mL PMA (P>.05), illustrating the ability of PMA‐qPCR to detect DNA exclusively from viable cells. Moreover, qPCR with 1.5 or 2.3 μg/mL PMA performed optimally with linearity over 10³‐10⁸ CFU/mL (R²≥0.9), whereas qPCR with 10, 23 or 46 μg/mL PMA significantly underestimated viable counts. Staphylococcus aureus and total viable bacteria were further determined with PMA‐qPCR (1.5 μg/mL) from 48 samples from a public library and two university dormitories and four from outside. Viable bacteria averaged 1.9×10⁴ cells/m³, and S. aureus were detected in 22 (42%) samples with a mean of 4.4×10³ cells/m³. The number of S. aureus and viable bacteria were positively correlated (r=.61, P<.005), and percentages of S. aureus relative to viable bacteria averaged 12‐44%. The results of field samples suggest that PMA‐qPCR can be used to quantify viable S. aureus cells.     

Thickening of waste activated sludge by biological flotation

Waste activated sludge was thickened by biological flotation without polymer flocculant dosage. The BIOFLOT® process utilizes the denitrifying ability of activated sludge bacteria. Gaseous products of anaerobic nitrate reduction cause spontaneous flotation of the sludge suspended solids. Laboratory tests confirmed the dependence of sludge thickening efficiency on available nitrate concentration, flotation time and temperature. Full-scale experiments were performed in a fully automatized unit for discontinuous sludge thickening from wastewater treatment plants with a capacity of up to 5000 I.E. Waste activated sludge from wastewater treatment plants at Pisek. Milevsko and Björnlunda was thickened from 6.2, 10.7 and 3.5 g/l MLSS to 59.4, 59.7 and 66.7 g/t MLSS, respectively. Concentrations of COD, ammonium and phosphate ions were decreased in sludge water. The average nitrate consumption for bioflotation was 21.2 mg NO₁⁻ per 1 g of MLSS of activated sludge. Flotation time ranged from 4 to 48 h.     

An examination of the surface characteristics of activated sludge in relation to bulking during the treatment of paper mill wastewater

The addition of mineral talc, Aquatal^®(Toulouse, France), to activated sludge treating paper mill effluents and its effect on settlement characteristics has been investigated. One laboratory study and a full‐scale investigation on a large waste water treatment plant (WWTP) with a capacity of 500.000 population equivalent (p.e.) have been carried out using this mineral. The sludge in the full‐scale plant was filamentous and had specific sludge volume index (SSVI) values in excess of 250. The talc dosing which was adapted to sludge settleability reduced SSVI values by 38%, improved the total suspended solids' concentrations by 86% and reduced the specific sludge load by 34%. In the laboratory‐scale plant, the sludge had SSVI values of 200–260 before treatment and was hydrophilic. The use of Aquatal^® at a dose of 0.7 g/g mixed liquor suspended solids (MLSS) took 2 weeks to reduce the SSVI to 78. It also increased the hydrophobicity to a balanced 50%. The use of mineral converted the somewhat diffuse flocs into a compact structure.